Genetically Diverse Mouse Embryonic Stem Cells: A Platform for Cellular Systems Genetics - PROJECT SUMMARY
The objective of this application is to generate a thoroughly-validated panel of genetically diverse mouse
embryonic stem cells (mESC) that will enable widespread adoption of cellular systems genetics. Phenotypic
variation, manifesting as heterogeneity in cell state, represents a significant challenge for realizing the full
promise of individualized, cell-based therapies, regenerative medicine. But phenotypic variation in genetically
diverse stem cells also presents an opportunity for the advancement of large scale, cellular screens of gene by
environment interactions (e.g. pharmacogenomics, toxicogenomics). A variety of approaches are beginning to
identify the networks that drive cell state transitions, but these efforts have largely focused on bulk assays,
which do not provide sufficient resolution of cell state heterogeneity, and mask the contribution of underlying
genetic variation on rare cell types. Moreover, genetic studies using human pluripotent stem cells are largely
limited to testing common variants due to low allele frequencies and imbalanced population structure requiring
prohibitively large samples and impeding identification of core regulatory networks with high power and
resolution. Therefore, we currently lack a thorough understanding of the genes and mechanisms that underlie
phenotypic variation in pluripotent stem cells. The Diversity Outbred (DO) mouse population at The Jackson
Laboratory is genetically defined, diverse, and presents a singular, cost-effective opportunity to systematically
investigate heterogeneity in mammalian pluripotency. Our pilot studies using DO mESCs establish the
feasibility of identifying regulatory loci at high power and resolution, as well as networks conserved in mice and
humans that regulate cell state transitions. In Aim 1, we will create a reference mapping panel of 300 DO
mESC lines that will serve as a gold standard resource for cellular systems genetics. This panel will be fully
credentialed and banked for broad availability through The Jackson Laboratory / Mutant Mouse Resource and
Research Centers (MMRRC). In Aim 2, we will determine at the single cell level the transcriptional networks
that regulate cell state transitions in vitro through the early stages of differentiation to mesoderm in a
representative subset of 144 lines. In Aim 3, we will map quantitative trait loci (QTL) that underlie variation in
cell state-specific gene expression and in the distribution of cell states in a population. In addition, we will build
and test models based on polygenic scores that can predict differentiation propensity from genotype. Finally, a
web-based searchable database of expression phenotypes and interactive tools for visualization of cell
composition and eQTL will be made publicly available to support community queries and hypothesis
generation. In sum, we will produce a resource of cell lines and gene expression data for the research
community that will spur new discoveries in regenerative medicine, pharmacogenomics, and toxicogenomics.
